rDsRed-Monomer Protein
rDsRed-Monomer Protein
Brand: Takara Bio.
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rDsRed-Monomer Protein
Affinity purified recombinant Discosoma sp. red fluorescent protein (rDsRed-Monomer) can be used as a standard for protein gels, Western blots, and fluorometry. It can also be used as a control for expression studies and for microinjection into cells and tissues. At its N-terminus, rDsRed-Monomer contains a 6xHN epitope tag, used for purification of the protein by immobilized metal affinity chromatography (IMAC) with our TALON Resin. An enterokinase (EK) site, located between the tag and rDsRed-Monomer, makes it possible to remove the 6xHN polypeptide if desired. The 6xHN tag does not interfere with the protein's characteristic spectral properties.
Overview
- Purified, recombinant proteins from E. coli
- Protein retains the absorption and emission spectra identical to expressed proteins
- Retains fluorescence capability under harsh conditions
Applications
- Western blotting
- Calibration of fluorescent protein detection instrumentation
- Microinjection in cells and tissues
DsRed-Monomer is soluble when expressed in mammalian cells
DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.
DsRed-Monomer is a monomeric protein
DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A280) and chromophore excitation (A557) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.
Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1
Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.
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DsRed-Monomer is soluble when expressed in mammalian cells
DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4% paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.
DsRed-Monomer is a monomeric protein
DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A280) and chromophore excitation (A557) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS PAGE analysis if samples are kept at 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) were separated by SDS PAGE electrophoresis (12% acrylamide). In both the boiled (denatured) and unboiled (nondenatured) samples, DsRed-Monomer fluorescent protein runs as a uniform band of ~30 kDa due to its monomeric structure. The unboiled (nondenatured) DsRed-Express runs at a much higher molecular weight than its boiled (denatured) counterpart due to its tetrameric structure.
Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1
Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.
